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UNROOTED STEM CUTTING PHYSIOLOGY; WATER USE and LEAF GAS EXCHANGE of SEVERED STEM CUTTINGS Peter Alem Clemson University, [email protected]

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UNROOTED STEM CUTTING PHYSIOLOGY; WATER USE and LEAF GAS EXCHANGE of SEVERED STEM CUTTINGS Peter Alem Clemson University, PALEM@CLEMSON.EDU Clemson University TigerPrints All Theses Theses 1-2010 UNROOTED STEM CUTTING PHYSIOLOGY; WATER USE AND LEAF GAS EXCHANGE OF SEVERED STEM CUTTINGS Peter Alem Clemson University, [email protected] Follow this and additional works at: https://tigerprints.clemson.edu/all_theses Part of the Horticulture Commons Recommended Citation Alem, Peter, "UNROOTED STEM CUTTING PHYSIOLOGY; WATER USE AND LEAF GAS EXCHANGE OF SEVERED STEM CUTTINGS" (2010). All Theses. 918. https://tigerprints.clemson.edu/all_theses/918 This Thesis is brought to you for free and open access by the Theses at TigerPrints. It has been accepted for inclusion in All Theses by an authorized administrator of TigerPrints. For more information, please contact [email protected]. UNROOTED STEM CUTTING PHYSIOLOGY; WATER USE AND LEAF GAS EXCHANGE OF SEVERED STEM CUTTINGS ___________________________________________________ A Thesis Presented To the Graduate School of Clemson University ____________________________________________________ In Partial Fulfillment of the Requirements for the Degree Master of Science Plant and Environmental Science _____________________________________________________ by Alem O. Peter August 2010 _____________________________________________________ Accepted by: Dr. Jim Faust, Committee Chair Dr. Christina Wells Dr. Dara Park Dr. Patrick Gerard ABSTRACT Stem cuttings are one of the most frequently used and successful methods of vegetative plant propagation. The understanding of unrooted cutting physiology, especially gas exchange and water flux, is crucial for successful propagation of healthy plants. Prior to root initiation, water uptake is limited and leafy stem cuttings are most vulnerable to wilting. Experiments were carried out in the greenhouse with poinsettia cuttings to determine water uptake, photosynthesis, transpiration and stomatal conductance from the time of severing to rooting, including time spent in storage. Water uptake through the severed stem was investigated in the laboratory and growth chamber. Cutting gas exchange (i.e., photosynthesis, stomatal conductance and transpiration) sharply declined after severing and gradually recovered in propagation after root initiation. CIRAS-2, with optional integrated Chlorophyll Fluorescence Module (CFM) was used to assess chlorophyll fluorescence and photosynthesis of cuttings at different stages: freshly severed cuttings (Day 1), 3 day-old cuttings, callused cuttings (Day 7 and 10), and rooted cuttings (Day 21 and 28). There was no significant difference in chlorophyll fluorescence between the cuttings and stock plants hence we inferred no significant damage to photosynthetic reaction centers as a result of severing. Unrooted cuttings had relatively low photosynthetic rates compared to rooted cuttings with water as a possible limiting factor. Our data also suggest that water use efficiency increases during the first week in propagation prior to root initiation. The mechanism for this improved water use appears to be due to a return in normal stomatal function during the first week in propagation ii following severing the cutting from the stock plant. The results have implications for the management of mist in propagation environment. iii ACKNOWLEDGEMENTS First, I would like to thank my advisor Dr. Jim Faust for his tireless support and help in helping me to understand and appreciate plant science and research and for offering an opportunity to be a student under his supervision. Secondly, I would like to thank Dr. Christina Wells for introducing and helping me learn how to use various instruments which were the means through which I gathered my data as well as for letting me use her laboratory and chemicals. I would also like to thank Dr. Dara Park who was instrumental in selecting appropriate courses for my degree, and Dr. Gerard who offered me advice on data analysis and report and accepted to be part of committee even though he was from another department. Finally, I would like to thank my brothers and sisters back in Kenya who are always there for me and Woollbrights family especially Joey and his family who have been great friends and a family to me. I would like to dedicate this work to my departed mother and father Akumu and Alem Limbe. iv TABLE OF CONTENTS TITLE PAGE ................................................................................................................... і ABSTRACT ....................................................................................................................ii ACKNOWLEDGEMENTS ........................................................................................... іiv TABLE OF CONTENTS ................................................................................................ v LIST OF TABLES ......................................................................................................... vi LIST OF FIGURES ....................................................................................................... vii CHAPTER 1. LITRATURE REVIEW ........................................................................................ 1 Water potential................................................................................................. 2 Stomatal behavior and functioning .................................................................. 4 Stomatal distribution ....................................................................................... 4 Factors responsible for opening and closing of stomata ................................... 5 Stomata and transpiration ................................................................................ 7 Factors affecting transpiration ......................................................................... 8 Water availability of rooting media ............................................................... 12 Relationship between photosynthesis and stomatal conductance .................... 13 Water movement in stem severed stems ........................................................ 14 Literature cited.. ............................................................................................ 16 2. WATER FLUX OF POINSETTIA CUTTINGS IN PROPAGATION ................ 23 Introduction .................................................................................................. 23 Materials and methods .................................................................................. 25 Results and Discussion .................................................................................. 28 Literature cited .............................................................................................. 38 3.GAS EXCHANGE OF POINSETTIA CUTTINGS (LEAVES) ......................... 39 Introduction .................................................................................................. 39 Materials and Methods .................................................................................. 42 Results and Discussion .................................................................................. 45 Literature cited .............................................................................................. 51 v LIST OF TABLES Table 2.1- Effect tests for initial water content, mist conditions (mist or no mist) and their interaction (Expt.2b)………………………..…………………………………...32 vi LIST OF FIGURES 1.1- Reflected images of stomata from intact leaves of Commelina communis and steady-state fluorescence imaging (Lawson, 2008…………………………………………………………7 2.1- Mean water loss during drought stress before wilting of unrooted cuttings…………………………………………………………………...………32 2.2- Mean time (min.) before wilting of unrooted cutting of 1, 3, 5 or 7 days old in propagation in the dark and in light (275 & 850 µmol m-2s-1)…………………………………………………...33 2.3- The effect of the initial water content of the cutting on the mean cumulative water uptake over 24……………………………………………………………..34 2.4- The effect of initial cutting water content on the rate of water uptake (ml/2h) (laboratory expt.) during the first 8 hours after placing the cutting in propagation………………………………………………………..35 2.5- The effect of initial cutting water content on the rate of water uptake (ml/2h) (under mist- greenhouse) during the first 8 hours after placing the cutting in propagation………………………………………………………..36 2.6- Mean cumulative water uptake through the stem of unrooted cuttings (1-7 days in propagation) during exposure to light (375 µmol m-2s-1) and in the dark for 48 h……………………………………….37 3.1- Gas exchange measurements included A) stomatal conductance, B) transpiration and C) photosynthesis from stock plant (Day 0) and cuttings (Days 1-3; in 10ºC cold, dark storage; Days 4-28 in propagation)……………………………………………………….48 3.2- Chlorophyll fluorescence (Fv/Fm) measured on stock plants (Day 0), and cuttings in propagation (Day1, 3, 5, 7, 21 or 28)………………………………………………………….48 3.3- Photosynthesis response curves generated by a chlorophyll fluorescence module mounted on the CIRAS-2 System……………………………………….50 vii CHAPTER ONE LITRATURE REVIEW Vegetative multiplication of plants is both a natural and an artificial process (Leakey, 1985). In nature, many plants by dividing through asexual structures to produce new plantlets for the next generation; they include corms, bulbils, runners and rhizomes, tuberous and bulbs (Bengtsson and Ceplitis, 2000). Artificial vegetative propagation of plants has been used for a long time and is still used by agriculturalists, horticulturalists, botanists and foresters, to propagate individual genotypes with preferred
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